The Leishmania cell surface metalloproteinase, leishmanolysin or GP63,
is expressed in all stages of Leishmania major. Initial studies repor
ted that in L. major the gp63 genes were arranged as five homologous,
tandemly repeated genes (gp63 genes 1-5) and a sixth, less conserved g
p63 gene located 8 kb downstream of gp63 gene 5. This study compared t
he sequences of L. major gp63 gene 1 and gp63 gene 6 and identified a
seventh L. major gp63 gene located downstream from gp63 gene 6. The L.
major gp63 genes exhibited stage-specific differences in their expres
sion: gp63 genes 1-5 were expressed in promastigotes only, gp63 gene 6
was expressed in promastigotes and amastigotes, while gp63 gene 7 was
expressed predominantly in stationary phase promastigotes and in amas
tigotes. Analysis of the predicted protein sequence of gp63 gene 6 (GP
63-6) and gp63 gene I (GP63-1) showed that these two proteins were hom
ologous in terms of overall predicted domain structure. L. major GP63-
1 has been reported to contain a glycosylphosphatidylinositol (GPI) me
mbrane anchor while sequence analysis predicted that GP63-6 contained
a different hydrophobic C-terminus that may act as a transmembrane reg
ion. Transfection studies using L. major gp63 gene 1 and gp63 gene 6 e
xpressed in L. donovani promastigotes showed that GP63-6 was expressed
at the cell surface and that the distinct GP63-6 C-terminus was capab
le of mediating GPI anchor attachment. (C) 1998 Elsevier Science B.V.
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